Fig. 6.

Gly-to-Asp substitution affects localization of NhhA in the outer membrane and trimer stability. (A) Western blot analysis of OMP preparations of the indicated M1390 and derivative strains subjected to denaturing conditions for subcellular detection of NhhA. Coomassie blue staining of the gel is shown in Fig. S4 in the supplemental material. Strain MC58 was used as a reference for trimeric localization of NhhA on OMPs. (B) Analysis of whole-cell lysates (WCL) from M1390 and its derivative strains prepared by alkaline lysis, subjected to seminative conditions, and analyzed by Western blotting. The Western blots were analyzed using serum anti-NhhA. The trimeric and monomeric forms are indicated by black and gray arrows, respectively. The asterisk marks a nonspecific cross-reactive band.